400mg leads to fatal intoxication in human

Inhibition of the human NET, DAT and SERT was assessed in HEK 293 cells that were stably transfected with the transporters. Cells were suspended in uptake buffer. We incubated the cells for 10 min with different concentrations of the test compounds (5-(2-Aminopropyl)benzofuran) and then added the corresponding [3H] monoamine (5 nM final concentration) at room temperature. After 10 min, we stopped uptake by separating the cells from the buffer using centrifugation through silicone oil. The centrifugation tubes were frozen in liquid nitrogen and cut to separate the cell pellet from the silicone oil and assay buffer layers. The cell pellet was then lysed. Scintillation fluid was added, and radioactivity was counted on a β-counter.